Accumulation of CD11c+CD163+ Adipose Tissue Macrophages through Upregulation of Intracellular 11 β-HSD1 in Human Obesity This information is current as Shotaro Nakajima, Vivien Koh, Ley-Fang Kua, Jimmy So, of September 24, 2021. Lomanto Davide, Kee Siang Lim, Sven Hans Petersen, Wei-Peng Yong, Asim Shabbir and Koji Kono J Immunol 2016; 197:3735-3745; Prepublished online 3 October 2016; doi: 10.4049/jimmunol.1600895 Downloaded from http://www.jimmunol.org/content/197/9/3735 Supplementary http://www.jimmunol.org/content/suppl/2016/10/01/jimmunol.160089 Material 5.DCSupplemental http://www.jimmunol.org/ References This article cites 62 articles, 14 of which you can access for free at: http://www.jimmunol.org/content/197/9/3735.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 24, 2021 • No Triage! 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The Journal of Immunology Accumulation of CD11c+CD163+ Adipose Tissue Macrophages through Upregulation of Intracellular 11b-HSD1 in Human Obesity Shotaro Nakajima,* Vivien Koh,† Ley-Fang Kua,† Jimmy So,‡ Lomanto Davide,‡ Kee Siang Lim,* Sven Hans Petersen,* Wei-Peng Yong,*,† Asim Shabbir,‡ and Koji Kono*,‡,x,{ Adipose tissue (AT) macrophages (ATMs) are key players for regulation of AT homeostasis and obesity-related metabolic disorders. However, the phenotypes of human ATMs and regulatory mechanisms of their polarization have not been clearly described. In this study, we investigated human ATMs in both abdominal visceral ATand s.c. ATand proposed an 11b-hydroxysteroid dehydrogenase type 1 (11b-HSD1)–glucocorticoid receptor regulatory axis that might dictate M1/M2 polarization in ATMs. The accumulation of Downloaded from CD11c+CD163+ ATMs in both visceral AT and s.c. AT of obese individuals was confirmed at the cellular level and was found to be clearly correlated with body mass index and production of reactive oxygen species. Using our in vitro system where human peripheral blood monocytes (hPBMs) were cocultured with Simpson–Golabi–Behmel syndrome adipocytes, M1/M2 polarization was found to be dependent on 11b-HSD1, an intracellular glucocorticoid reactivating enzyme. Exposure of hPBMs to cortisol- induced expression of CD163 and RU-486, a glucocorticoid receptor antagonist, significantly abrogated CD163 expression through coculture of mature adipocytes with hPBMs. Moreover, 11b-HSD1 was expressed in crown ATMs in obese AT. Importantly, http://www.jimmunol.org/ conditioned medium from coculture of adipocytes with hPBMs enhanced proliferation of human breast cancer MCF7 and MDA- MB-231 cells. In summary, the phenotypic switch of ATMs from M2 to mixed M1/M2 phenotype occurred through differentiation of adipocytes in obese individuals, and upregulation of intracellular 11b-HSD1 might play a role in the process. The Journal of Immunology, 2016, 197: 3735–3745. verweight and obese individuals pose global health is- have been postulated as important elements in inflammation (4, 5). sues and are associated with increased risks of metabolic Understanding the regulatory mechanisms of ATM polarization O diseases, including type 2 diabetes, cardiovascular dis- can be important in preventing the development of obesity-related by guest on September 24, 2021 ease, and certain cancers (1–3). Obesity and related metabolic metabolic disorders. However, human ATM phenotypes and their disorders are associated with a state of chronic, low-grade in- polarization mechanisms remain unclear. flammation in adipose tissue (AT), and AT macrophages (ATMs) There are at least two different phenotypes of ATMs: classically activated proinflammatory ATMs (M1 ATMs) and alternatively activated anti-inflammatory ATMs (M2 ATMs). In nonobese ani- *Center for Translational Medicine, Cancer Science Institute of Singapore, National Uni- versity of Singapore, Singapore 117599; †Department of Hematology–Oncology, National mals, resident M2 ATMs are predominantly observed and con- University of Singapore, Singapore 119228; ‡Department of Surgery, National University tribute to maintenance of AT homeostasis. Alternatively, during of Singapore, Singapore 119228; xDepartment of Organ Regulatory Surgery, Fukushima Medical University, Fukushima 960-1295, Japan; and {Department of Advanced Cancer obesity, secretions of chemotactic molecules, such as MCP-1 from Immunotherapy, Fukushima Medical University, Fukushima 960-1295, Japan hypertrophic adipocytes, result in the recruitment of circulating ORCID: 0000-0001-9330-5783 (K.S.L.). monocytes into AT and their differentiation into an M1 phenotype Received for publication May 24, 2016. Accepted for publication September 1, 2016. (6, 7). Indeed, this phenotypic switch from M2 to M1 in AT is a This work was supported by the National Medical Research Council of Singapore crucial determinant of insulin resistance in obese mice. In mice (the Clinician Scientist Award and the Singapore Translational Research Investigator fed a high-fat diet, a marked increase in the accumulation of F4/ Award) (to K.K.). 80+CD11c+ ATM–expressing inflammatory genes, as well as the The microarray data presented in this article have been submitted to the Gene Ex- depletion of these F4/80+CD11c+ cells, would normalize obesity- pression Omnibus (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE86492) under accession number GSE86492. induced insulin resistance (8, 9). In contrast, other groups reported Address correspondence and reprint requests to Prof. Koji Kono, Department of Organ that this phenotypic switch in human ATMs in obese individuals is Regulatory Surgery and Department of Advanced Cancer Immunotherapy, Fukushima fundamentally different from the mouse model. Zeyda et al. (10) Medical University, Fukushima 960-1295, Japan. E-mail address: [email protected] and Bourlier et al. (11) reported that human ATMs were of the M2 The online version of this article contains supplemental material. phenotype–producing proinflammatory mediators. Another study Abbreviations used in this article: AT, adipose tissue; ATM, AT macrophage; BMI, by Wentworth et al. (12) suggested that the percentage of CD11c+ body mass index; CLS, crown-like structure; CM, conditioned medium; ER, endo- + plasmic reticulum; FFA, free fatty acid; GR, glucocorticoid receptor; HbA1c, hemo- CD206 ATMs with mixed M1/M2 phenotype increased in obese globin A1c; hPBM, human peripheral blood monocyte; 11-bHSD, 11b-hydroxysteroid subjects and was correlated with markers of increased insulin dehydrogenase; 11b-HSD1, 11b-hydroxysteroid dehydrogenase type 1; 11b-HSD2, 11b-hydroxysteroid dehydrogenase type 2; qPCR, quantitative real-time PCR; ROS, resistance, such as crown-like structures (CLS), and also with the reactive oxygen species; SAT, s.c. AT; SGBS, Simpson–Golabi–Behmel syndrome; homeostasis model assessment of insulin resistance. Importantly, SVF, stromal vascular fraction; TAM, tumor-associated macrophage; VAT, visceral Becker and colleagues (13) demonstrated that metabolic activation AT; VEGF, vascular endothelial growth factor. pathways triggered by glucose, insulin, and palmitate also contribute Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 to ATM-specific polarization in humans and mice. Therefore, the www.jimmunol.org/cgi/doi/10.4049/jimmunol.1600895 3736 INCREASING CD11c+CD163+ ATMs IN HUMAN OBESITY regulation of ATM polarization is complex, and little is known of the human CD68 Ab (Dako, Glostrup, Denmark); human 11b-HSD1 Ab link between human ATM phenotype and metabolic disorders in (Abcam, Cambridge, U.K.); and b-actin Ab (Cell Signaling Technology, obese individuals. Further investigations on the phenotype of human Danvers, MA). ATMs and their functions are needed to develop efficient therapies Collection of AT for obesity and related metabolic diseases. This study was approved by the Domain-Specific Ethics Board of the Several markers of human M1 and M2 ATMs have been National University Hospital of Singapore (DSRB no. 2015/00031). We reported. Cell surface markers of M1 ATMs include CD11c, CD40, recruited 7 nonobese subjects (BMI , 30 kg/m2) and 22 obese subjects CD86, HLA-DR, and TLR4 (7, 14, 15). CD11c is predominantly (BMI $ 30 kg/m2) from the National University Hospital of Singapore expressed by crown ATMs in the proinflammatory state (12). (Table I). Abdominal VAT (omental AT) and SAT were collected during herniorrhaphy for nonobese subjects and during bariatric surgery for obese Several groups also reported CD11c as a human M1 marker subjects after obtaining informed consent. None of the subjects had any (16–18). Thus, it is considered to be a suitable marker of human clinical symptoms of systemic inflammation. M1 ATMs. Additionally, there are several known cell surface Isolation of stromal vascular fraction from AT